Construction and characterization of hGM-CSF-expressing K-562 cell line
- Author:
Yinxia CHEN
;
Suhu LIU
;
Yun YANG
;
Jie LIU
;
Xiaorong MA
;
Wanggang ZHANG
- Publication Type:Journal Article
- Keywords:
K-562 cell;
GM-CSF;
vaccine
- From:
Journal of Pharmaceutical Analysis
2007;19(2):200-203
- CountryChina
- Language:Chinese
-
Abstract:
Objective The whole process of vaccine preparation is time-consuming and technically challenging. Here the hGM-CSF-engineered K-562 cell line was constructed to simplify tumor vaccine preparation process. Methods The eukaryocyte expressing plasmid pcDNA3.1/GM-CSF was first constructed and its accuracy was verified through sequencing. The pcDNA3.1/GM-CSF was transfected into COS-7 cells to verify GM-CSF expression and cytokine activity using TF-1 cell line. Then the plasmid was transfected into K-562 cell line using liposome method, and was selected under G-418 and sub-cloned by limiting dilution. GM-CSF product from the monoclone GM-CSF-K-562 cell lines was quantified using ELISA method. Results We successfully constructed the hGM-CSF eukaryocyte expressing plasmid and hGM-CSF expressing K-562 cell line. Conclusion The construction of K-562/GM-CSF line will simplify the preparation of tumor vaccine, thus facilitating the application of tumor vaccination therapy in clinical application.
